Three-dimensional imaging of whole rodent organs using optical computed and emission tomography

We explore the potential of optical computed tomography (optical-CT) and optical emission computed tomography (optical-ECT) in a new area-whole organ imaging. The techniques are implemented on an in-house prototype benchtop system with improved image quality and the capacity to image larger samples (up to 3 cm) than previous systems based on stereo microscopes. Imaging performance tests confirm high geometrical accuracy, accurate relative measurement of linear attenuation coefficients, and the ability to image features at the 50-microm level. Optical labeling of organ microvasculature was achieved using two stains deposited via natural in vivo circulatory processes: a passive absorbing ink-based stain and an active fluorescin FITC-lectin conjugate. The lectin protein binds to the endothelial lining, and FITC fluorescense enables optical-ECT imaging. Three-dimensional (3-D) optical-CT images have been acquired of a normal rat heart and left lung and a mouse right lung showing exquisite detail of the functional vasculature and relative perfusion distribution. Coregistered optical-ECT images were also acquired of the mouse lung and kidney. Histological sections confirmed effective labeling of microvasculature throughout the organs. The advantages of optical-CT and optical-ECT include the potential for a unique combination of high resolution and high contrast and compatibility with a wide variety of optical probes, including gene expression labeling fluorescent reporter proteins.     

Three-dimensional cellular-level imaging using full-field optical coherence tomography

An ultrahigh-resolution full-field optical coherence tomography (OCT) system has been developed for cellular-level imaging of biological media. The system is based on a Linnik interference microscope illuminated with a tungsten halogen lamp, associated with a high-resolution CCD camera. En face tomographic images are produced in real time, with the best spatial resolution ever achieved in OCT (0.7 microm x 0.9 microm, axial x transverse). A shot-noise limited detection sensitivity of 80 dB can be reached with an acquisition time per image of 1 s. Images of animal ophthalmic biopsies and vegetal tissues are shown.     

Three-dimensional imaging and osteometry of adult human skulls using helical computed tomography

To make a digital image database of human craniology, we optimized the three-dimensional (3-D) images of 29 dried human skull specimens by helical computed tomography (CT). For the verification of the quantitative exactitude of these image data, we manually measured nine items of direct distances between standard anthropologic points on each skull and the corresponding distances projected on the CT monitor by specifying the respective points. The results obtained by the two methods of manual and CT measurements were compared and statistically analyzed. The CT measurements were so exact that the lower limit of correlation coefficients (95% of the confidence interval) between the two results was more than 0.8 in six items; i.e., maximal cranial length and breadth, minimal frontal breadth, bizygomatic breadth, distance between ectomolares and nasion-basion length. In contrast, the CT results were less well correlated with the manual measurements of three items; i.e., distance between bilateral mastoidales, total facial height, and nasal breadth. We concluded that the qualitative representation of 3-D CT images was adequate, although some quantitative data may be incorrect. The inaccuracy is suspected to be due to the difficulty in specifying the standard points on the CT images, and due to the differences in measurement procedures between the direct and projected distances.     

Three-dimensional fluorescence-enhanced optical tomography using a hand-held probe based imaging system

Hand-held based optical imaging systems are a recent development towards diagnostic imaging of breast cancer. To date, all the hand-held based optical imagers are used to perform only surface mapping and target localization, but are not capable of demonstrating tomographic imaging. Herein, a novel hand-held probe based optical imager is developed towards three-dimensional (3-D) optical tomography studies. The unique features of this optical imager, which primarily consists of a hand-held probe and an intensified charge coupled device detector, are its ability to; (i) image large tissue areas (5 x 10 sq. cm) in a single scan, (ii) perform simultaneous multiple point illumination and collection, thus reducing the overall imaging time; and (iii) adapt to varying tissue curvatures, from a flexible probe head design. Experimental studies are performed in the frequency domain on large slab phantoms (approximately 650 ml) using fluorescence target(s) under perfect uptake (1:0) contrast ratios, and varying target depths (1-2 cm) and X-Y locations. The effect of implementing simultaneous over sequential multiple point illumination towards 3-D tomography is experimentally demonstrated. The feasibility of 3-D optical tomography studies has been demonstrated for the first time using a hand-held based optical imager. Preliminary fluorescence-enhanced optical tomography studies are able to reconstruct 0.45 ml target(s) located at different target depths (1-2 cm). However, the depth recovery was limited as the actual target depth increased, since only reflectance measurements were acquired. Extensive tomography studies are currently carried out to determine the resolution and performance limits of the imager on flat and curved phantoms.     

In-situ imaging of articular cartilage of the first carpometacarpal joint using co-registered optical coherence tomography and computed tomography

Conventional imaging modalities are unable to depict the early degeneration of articular cartilage in osteoarthritis, especially in small joints. Optical coherence tomography has previously been used successfully in high-resolution imaging of cartilage tissue. This pilot cadaver study demonstrates the use of intra-articular optical coherence tomography in imaging of articular cartilage of the first carpometacarpal joint, producing high resolution images of the articular surface in which cartilage thickness and surface characteristics were assessed. Findings on optical coherence tomography were confirmed with histology. Furthermore, co-registration of optical coherence tomography and computed tomography was used to accurately determine the scanned trajectory and reconstruct a true-scale image overlay.     

Three-dimensional computed tomography imaging in the evaluation of craniofacial abnormalities.

A retrospective study was undertaken to assess the value of three-dimensional computed tomography (3-D CT) in evaluating craniofacial abnormalities. Over a 2-year period, 145 children evaluated with routine two-dimensional (2-D) CT had 3-D CT reformatted images assessing a variety of anomalies including craniosynostosis, midface clefts, trauma, and craniofacial syndromes. There were 62 females and 83 males ranging in age from 1 month to 17 years (median: 5 months). Both soft-tissue and bone algorithms were used. In all 145 cases, the 3-D CT images confirmed or provided additional information of depth perception, contours, volumes, and extent of an abnormality. This was especially important and useful in the settings of complex anomalies and preoperative planning. We concluded that 3-D CT reconstructed images in conjunction with routine 2-D CT should be an integral part of the examination in evaluating craniofacial abnormalities.     

Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate or BABB. The optimized optical clearing procedure of ethanol fixation followed by methyl salicylate clearing preserved the fluorescence of constitutive RFP in whole xenograft tumour specimens, about 1 cc in dimension, indicating successful extension from cell plating experiments to whole tissue samples. Finally, the feasibility of imaging the 3D distribution of viable tumour cells (as indicated by the RFP emission) is demonstrated by optical-ECT imaging of cleared xenograft tumours using an in-house system.     

Single-photon emission computed tomography imaging for brain death donor counseling

Organ donation awareness is very poor in India. We have a high demand for transplant organs with poor supply. Apnea test is the confirmatory test for brain death in our country. The Transplantation of Human Organs Act does not support any ancillary testing for the confirmation of brain death in our country. Radionuclide scan is used widely in western countries as a confirmatory test. We in our institution used this as a tool for family counseling with successful conversion rate.Key words: Organ donation, radionuclide imaging, single-photon emission computed tomography     

正电子发射计算机断层显像/核磁共振：分子影像学技术新进展

背景：正电子发射计算机断层显像(PET)从分子水平反映细胞代谢和功能变化,MRI有更高的软组织对比度和空间分辨率,且无电磁辐射。近年来,随着多模式成像技术飞速发展,作为刚刚应用于临床的多模式分子成像技术,PET/MRI融合显像可提供分子、形态与功能信息,有关研究已成为分子影像领域的关注焦点。 目的：就目前PET/MRI研制存在的问题和进展情况做一综述,并展望其潜在的临床和科研价值。 方法：由第一作者应用计算机检索CNKI数据库、Springerlink数据库、Pubmed数据库相关文献。检索时间范围：2000年1月至2012年6月。英文检索词：“PET/MRI”和“multimodality imaging or image fusion”,中文检索词：“分子影像学”和“图像融合”。选择内容与PET/MRI多模式成像相关的文章,同一领域文献则选择近期发表或发表在权威杂志文章,共纳入53篇文献。 结果与结论：已有的研究成果表明,PET/MRI较传统显像手段和其他成功的多模式成像技术,如PET/CT和SPECT/CT有更多、更新的应用优势,并已积累临床前和初步临床应用的经验,PET/MRI具有广阔的发展空间,有望将分子影像技术的发展带入一个新的时代,为临床诊断掀开新的篇章。     

Three-dimensional analysis of deciduous maxillary anterior teeth using cone-beam computed tomography

The recent introduction of cone-beam computed tomography (CBCT) into the medical field has allowed the nondestructive investigation of internal structures at relatively low cost and radiation exposure. The accuracy of CBCT in both two and three dimensions has been demonstrated, and CBCT has been used successfully for craniofacial anatomy. Knowing the anatomical structure of deciduous teeth is essential for clinical dentistry. However, the root structure of deciduous teeth is rarely reported because of the scarcity of intact deciduous teeth without root resorption. The aim of this study was to evaluate the intact root form of deciduous teeth using CBCT. Data from 38 young children was analyzed using an image-analyzing program. The degree of buccal dilacerations was 26.3° for deciduous maxillary central incisors (DMA), 16.5° for deciduous maxillary lateral incisors (DMB), and 17.5° for deciduous maxillary canines (DMC) in about half of the root length. The crown-to-root ratios were 0.52 for DMA, 0.48 for DMB, and 0.52 for DMC. These data will be helpful for understanding the development of dentition, and for clinical dentistry.     

Cone-beam optical computed tomography for gel dosimetry II: imaging protocols

This work develops imaging protocols for improved dose readout of a Fricke-xylenol orange-gelatin (FXG) gel-filled 1 L polyethylene terephthalate (PETE) jar dosimeter using a commercial Vista(TM) cone-beam optical computed tomography (CT) scanner from Modus Medical Devices Inc. (London, ON, Canada). To ensure good management of light source-detector stability, it was determined that (a) a minimum of 2 h warm-up time is necessary prior to dosimeter scanning, (b) the light source should be kept on until the completion of the last data scan except for the minimum amount of time required to acquire dark field images, and (c) the optional Vista software projection image normalization routine should be used in image reconstruction. The institution of dosimeter scan time and temperature control was strongly indicated from the experiments. A standard post-irradiation wait time of 30 min measured to within ±30 s was established to minimize the measurement uncertainties due to dosimeter development and diffusion. To alleviate thermochromic behavior leading to inaccurate dose readout, holding bath warm up and pre-scan temperature adjustment procedures were developed to control dosimeter temperature to within ±0.2 °C. The possibility of stray light minimizing protocols was also investigated and deemed to be unnecessary. The largest significant sources of stray light in the system were identified as being due to angled scatter from the dosimeter gelatin matrix and refraction from the jar wall interfaces. It was concluded that these phenomena would be better addressed through dosimeter modification and an inter-jar dose-to-attenuation calibration methodology, rather than by setting additional imaging protocols.     

Non-invasive imaging of adaptive immunity using positron emission tomography

Summary: Non-invasive monitoring of adaptive immunity in infection, cancer, and autoimmunity remains a major challenge. Current techniques to monitor lymphocytes involve numeric and functional determinations of immune cells isolated from the peripheral blood (most often) and tissue (rarely). Invasive measurements are prone to sampling errors and are poorly reflective of the dynamic changes in the location, number, and movement of lymphoid cells. These limitations indicate the need for non-invasive whole-body imaging methodologies that allow longitudinal, quantitative, and functional analyses of the immune system in vivo . Positron emission tomography (PET), a clinically based whole-body imaging modality, has the potential to revolutionize diagnostics and therapeutic monitoring in both clinical and pre-clinical settings. This review discusses studies using PET to image adaptive immune responses in small animal models. We address the challenges inherent in assessing whole-body immunity with PET and recent developments that can improve its performance. Finally, we discuss work to translate PET immune imaging into clinical practice.     

基于4D-CT最大呼吸气相位图像获取三维肺通气的信息

背景：传统的肺功能成像技术存在诸多不便,利用4D-CT中蕴含的通气信息进行功能图像的快速提取对肺部疾病的诊断和治疗有非常重要的意义。 目的：探讨基于三维变形图像配准算法从4D-CT最大吸气相位和最大呼气相位图像中获取肺通气的三维分布的可行性。 方法：利用电影模式采集自由呼吸状态下的胸部CT图像并利用已开发的4D-CT软件进行四维重建,得到吸气末和呼气末双相位CT图像,依次进行肺组织分割、利用基于体积的变形图像配准算法进行三维图像配准、量化分析三维空间象素的位移矢量,最后得到通气度量图即肺功能区的三维分布图。 结果与结论：利用三维变形图像配准算法,实现了从4D-CT最大吸气相位和最大呼气相位图像中获取在任意横断位、冠状位和矢状位的肺通气分布。     

Direct reconstruction of single-photon emission computed tomography images using retained matrix elements

In clinical applications, two methods of single-photon emission computed tomography (SPECT) reconstruction are widely used. These are filtered backprojection and iterative reconstruction. Filtered backprojection is fast and produces acceptable images. Iterative reconstruction is slow, but produces images of greater accuracy than backprojection. The authors sought to develop a method of SPECT reconstruction that would have the advantages of both established methods: close in speed to backprojection and with the accuracy of iterative reconstruction. This was accomplished by computing a direct solution to the set of linear equations governing SPECT reconstruction. We tested this method of SPECT reconstruction using a set of projections from a cold rod and sphere phantom. Direct reconstruction produced images having equivalent resolution to backprojected images, but with double the contrast ratio. The direct method required 10 seconds of computation per slice on a Macintosh Quadra 950 (Apple Computer; Cupertin, CA), significantly faster than most iterative methods.     

Three-dimensional diffuse optical tomography of bones and joints

We present for the first time full three-dimensional (3D) volumetric reconstruction of absorption images of in vitro and in vivo bones and joints from near-infrared tomographic measurements. Imaging experiments were conducted on human finger and chicken bones embedded in cylindrical scattering media using a Clemson multichannel diffuse optical imager. The volumetric optical images were recovered with our 3D finite element based reconstruction algorithm. Our results show that 3D imaging methods can provide details of the joint structure/composition that would be impossible from two-dimensional imaging methods.     

Optical clearing of unsectioned specimens for three-dimensional imaging via optical transmission and emission tomography

Optical computed tomography (optical-CT) and optical emission computed tomography (optical-ECT) are new techniques that enable unprecedented high-resolution 3-D multimodal imaging of tissue structure and function. Applications include imaging macroscopic gene expression and microvasculature structure in unsectioned biological specimens up to 8 cm(3). A key requisite for these imaging techniques is effective sample preparation including optical clearing, which enables light transport through the sample while preserving the signal (either light absorbing stain or fluorescent proteins) in representative form. We review recent developments in optical-CT and optical-ECT, and compatible "fluorescence-friendly" optical clearing protocols.